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Construction and Characterization of a Bacterial Artificial Chromosome (BAC) Library of Pacific White Shrimp, Litopenaeus vannamei

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Abstract

The pacific white shrimp (Litopenaeus vannamei) is one of the most economically important marine aquaculture species in the world. To facilitate gene cloning and characterization, genome analysis, physical mapping, and molecular selection breeding of marine shrimp, we have developed the techniques to isolate high-quality megabase-sized DNA from hemocyte nuclear DNA of female shrimp and constructed a bacterial artificial chromosome (BAC) genomic library for the species. The library was constructed in the Hind III site of the vector pECBAC1, consisting of 101,760 clones arrayed in 265 384-well microtiter plates, with an average insert size of 101 kb, and covering the genome approximately fivefold. To characterize the library, 92,160 clones were spotted onto high-density nylon filters for hybridization screening. A set of 18 pairs of overgo probes designed from eight cDNA sequences of L. vannamei genes were used in hybridization screening, and 35 positive clones were identified. These results suggest that the shrimp BAC libraries will provide a useful resource for screening of genomic regions of interest candidate genes, gene families, or large-sized synthetic DNA region and promote future works on comparative genomics, physical mapping, and large-scale genome sequencing in the species.

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Acknowledgments

This project is supported by National High-Tech Research and Development Program of China (2007AA09Z430), Key Program of National Natural Science Foundation of China (No. 30730071), Major State Basic Research Development Program of China (2006CB101804) and Public Industry (Agriculture) Specific Research Program (No.200803012). The authors thank Prof. Linsheng Song, Dr. Baozhong Liu, Dr. Jiquan Zhang, Hao Jiang, Cui Zhao, Fan He, and other laboratory members for technical advice and assistance.

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Authors and Affiliations

  1. Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, China

    Xiaojun Zhang, Yang Zhang, Pin Huan, Bing Wang, Chengzhang Liu, Fuhua Li, Bin Liu & Jianhai Xiang

  2. Graduate University of the Chinese Academy of Sciences, Beijing, 100039, China

    Yang Zhang & Pin Huan

  3. Department of Soil & Crop Sciences, Texas A&M University, College Station, TX, 77843-2474, USA

    Yang Zhang, Chantel Scheuring & Hong-Bin Zhang

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  1. Xiaojun Zhang
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  2. Yang Zhang
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  3. Chantel Scheuring
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  4. Hong-Bin Zhang
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  5. Pin Huan
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  6. Bing Wang
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  7. Chengzhang Liu
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  8. Fuhua Li
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  9. Bin Liu
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  10. Jianhai Xiang
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Correspondence to Jianhai Xiang.

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Xiaojun Zhang and Yang Zhang contributed equally to the article.

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Zhang, X., Zhang, Y., Scheuring, C. et al. Construction and Characterization of a Bacterial Artificial Chromosome (BAC) Library of Pacific White Shrimp, Litopenaeus vannamei . Mar Biotechnol 12, 141–149 (2010). https://doi.org/10.1007/s10126-009-9209-y

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